A highly sensitive and specific real-time quantitative polymerase chain reaction assay for Perkinsus marinus detection in oysters.

Sci Rep

Department of Aquatic Life Medicine, College of Ocean Science and Technology, Kunsan National University, 558 Daehakro, Gunsan, 54150, South Korea.

Published: October 2024

Oyster aquaculture is one of the fastest-growing food production industries worldwide; however, it faces a significant challenge from the protist Perkinsus marinus, particularly in the USA. Although several quantitative molecular diagnostic methodologies are available for identifying diseases caused by P. marinus, the primer pairs used therein led to non-specific identification of other Perkinsus spp. Hence, a quantitative real-time PCR (Pm-qPCR) assay specific for P. marinus was developed using a TaqMan-based probe with the internal quencher in this study. A primer pair and probe specific to P. marinus were designed from a hypothetical protein of P. marinus collected from the whole-genome shotgun sequence database of the National Center for Biotechnology Information (NCBI). In silico analysis using homologous sequences of P. olseni and P. chesapeaki confirmed the high specificity of primers designed in this study. The Pm-qPCR assay was performed using seven different strains of P. marinus, P. olseni, and P. chesapeaki, revealing high specificity and sensitivity for detecting only P. marinus strains. In conclusion, it was demonstrated that Pm-qPCR can effectively and accurately diagnose P. marinus with high specificity and sensitivity. This assay is promising for monitoring oyster health and disease management in ecosystems and aquaculture.

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC11513098PMC
http://dx.doi.org/10.1038/s41598-024-76822-yDOI Listing

Publication Analysis

Top Keywords

high specificity
12
marinus
9
perkinsus marinus
8
pm-qpcr assay
8
specific marinus
8
olseni chesapeaki
8
specificity sensitivity
8
highly sensitive
4
sensitive specific
4
specific real-time
4

Similar Publications

Want AI Summaries of new PubMed Abstracts delivered to your In-box?

Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!